| In this paper, Palygorskite(PGS) was used as the carrier of ginger essential oil (GEO) and a novel kind of composite GEO-PGS was prepared, which was optimized by single-factor test and the response surface method (RSM). The properties of GEO-PGS were characterized by FTIR, XPS, XRD, TG and SEM. With the use of tests in vivo and in vitro, we can do research GEO-PGS inhibition against Staphylococcus aureus, and study its mechanism of inhibition from the molecular level. The following is the main content and result:1、Preparation of GEO-PGS:PGS was used as the carrier of GEO and a novel kind of composite GEO-PGS was prepared in our laboratory, which was optimized by single-factor test and the response surface method (RSM). The optimal preparation conditions of GEO-PGS composite were the reaction time 150.44min, initial [6]-gingerol concentration of GEO 214.11mg·mL-1, and the reaction temperature 80.56℃. [6]-gingerol-loaded amount of GEO-PGS composite was 0.93763% by HPLC analysis, which was in accordance with the results predicted by RSM model.2、Characterization of GEO-PGS:The properties of GEO-PGS were characterized by FTIR, XPS, XRD, TG and SEM. Results of FTIR and XPS analysis demonstrated the combination of GEO and PGS. XRD analysis showed that GEO was absorbed on the surface of PGS. TG analysis confirmed the combination of GEO and PGS, and the content of GEO in GEO-PGS being estimated about 18.66%. SEM observation showed that the morphology of GEO-PGS changed obviously compared with that of PGS, which also demonstrated the combination of GEO and PGS. Dispersion analysis showed GEO-PGS realized organic and its nature has changed, GEO-PGS has significant hydrophobic and lipophilic.3、In vitro antibacterial activity of GEO-PGS:In this paper, we studied the antibacterial effect of GEO-PGS by both in vitro and in vivo. Use the two-fold dilution method to measure the minimum inhibitory concentration (MIC) of Staphylococcus aureus, the MIC was 3.5625 mg·ml-1 and after PGS was used as the carrier of GEO, GEO has a stronger antibacterial activity. After extended period of bacterial growth and the logarithm of Joining GEO-PGS, bacterial growth is inhibited. Gram staining test showed that GEO-PGS had the specific-targeting antibacterial capability. The novel composite GEO-PGS combined the bacteria-absorbent activity of PGS and the antibacterial activity of GEO. PGS was used as the carrier of GEO, GEO showed the characteristics of thermo-stability, acidity-resistance, alkalinity-resistance, Weather-resistance and aging-resistance.In vivo antibacterial activity of GEO-PGS:By subcutaneous injection of Staphylococcus aureus in Kunming mice in vivo, establish an animal model of skin infection. Set six groups, blank group, model group, positive control group (Bactroban), Ginger group, GEO-PGS low-dose group, GEO-PGS high-dose group, continuous smear 7 days. The results showed that model mice with smear GEO-PGS, the lesion area was significantly reduced, the skin scores were better than the model group and ginger group, lesions were significantly improved conditions, Weight gradually restored, arranged nucleus tends structured, inflammatory cell infiltration is improved and the expansion of the capillary phenomenon slowed. All these description that GEO-PGS can better treat bacterial infections of the skin ulcers and inflamed repaired. With respect to the GEO, PGS has a better therapeutic effect and shorter onset time.4、Molecular mechanism of action of the Antibacterial Activity:Detected the affect of GEO-PGS treatment on the skin infection model related gene expression. Genes involved have Toll-like receptor 2 (TLR-2), Toll-like receptor 4 (TLR-4), interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α). The results showed that Toll--like receptor 2 (TLR-2), Toll-like receptor 4 (TLR-4), interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α) gene expression in the administered group was significantly lower than the model group, but higher than the blank control and positive control group. Description GEO-PGS has a role in reduced Staphylococcus aureus infection inflammatory cytokines and induced innate immune system may be a good treatment of the Staphylococcus aureus infections of the skin inflammation.In summary, GEO-PGS was prepared in our laboratory. GEO-PGS combined the bacteria-absorbent activity of PGS and the antibacterial activity of GEO. Moreover, GEO-PGS showed the characteristics of thermo-stability, acidity-resistance, alkalinity-resistance, Weather-resistance and aging-resistance. GEO-PGS has a good inhibition effect on Staphylococcus aureus and to treat skin inflammation due to the bacterial infection, suggesting the great potential application of GEO-PGS as the novel antibacterial composite substance. |
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